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      Understanding Stress Response to High-Arsenic Gold-Bearing Sulfide Concentrate in Extremely Metal-Resistant Acidophile Sulfobacillus thermotolerans

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          Abstract

          Biooxidation of gold-bearing arsenopyrite concentrates, using acidophilic microbial communities, is among the largest commercial biohydrometallurgical processes. However, molecular mechanisms of microbial responses to sulfide raw materials have not been widely studied. The goal of this research was to gain insight into the defense strategies of the acidophilic bacterium Sulfobacillus thermotolerans, which dominates microbial communities functioning in industrial biooxidation processes at >35 °C, against the toxic effect of the high-arsenic gold-bearing sulfide concentrate. In addition to extreme metal resistance, this acidophile proved to be one of the most As-tolerant microorganisms. Comparative proteomic analysis indicated that 30 out of 33 differentially expressed proteins were upregulated in response to the ore concentrate, while the synthesis level of the functional proteins required for cell survival was not negatively affected. Despite a high level of cellular metal(loid) accumulation, no specific metal(loid)-resistant systems were regulated. Instead, several proteins involved in the metabolic pathways and stress response, including MBL fold metallo-hydrolase, sulfide:quinone oxidoreductase, and GroEL chaperonin, may play crucial roles in resistance to the sulfide ore concentrate and arsenic, in particular. This study provides the first data on the microbial responses to sulfide ore concentrates and advances our understanding of defense mechanisms against toxic compounds in acidophiles.

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          Most cited references81

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          Clp ATPases are required for stress tolerance, intracellular replication and biofilm formation in Staphylococcus aureus.

          The Hsp100/Clp ATPases constitute a family of closely related proteins of which some members function solely as chaperones whereas others additionally can associate with the unrelated ClpP peptidase forming a Clp proteolytic complex. We have investigated the role of four Clp ATPases in the versatile pathogen, Staphylococcus aureus. Previously, we showed that ClpX is required for expression of major virulence factors and for virulence of S. aureus, but not for survival during heat shock. In the present study, we have inactivated clpC, clpB and clpL and, while none of these mutations affected toxin production, both ClpC and ClpB and to a minor extent ClpL were required for intracellular multiplication within bovine mammary epithelial cells. These defects were paralleled by an inability of the clpC mutant to grow at high temperature and of the clpB mutant to induce thermotolerance indicating that the protective functions of these proteins are required both at high temperature and during infection. By primer extension analysis and footprint studies, we show that expression of clpC and clpB is controlled by the negative heat-shock regulator, CtsR, and that ClpC is required for its repressor activity. Thus, ClpC is a likely sensor of stress encountered during both environmental stress and infection. In addition to virulence factor production the ability to form biofilms is of importance to S. aureus as a nosocomial pathogen. Interestingly, biofilm formation was reduced in the absence of ClpX or ClpC whereas it was enhanced in the absence of ClpP. Thus, our data show that Clp proteolytic complexes and the Clp ATPases control several key processes of importance to the success of S. aureus as a pathogen.
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            The microbiology of biomining: development and optimization of mineral-oxidizing microbial consortia.

            Biomining, the use of micro-organisms to recover precious and base metals from mineral ores and concentrates, has developed into a successful and expanding area of biotechnology. While careful considerations are made in the design and engineering of biomining operations, microbiological aspects have been subjected to far less scrutiny and control. Biomining processes employ microbial consortia that are dominated by acidophilic, autotrophic iron- or sulfur-oxidizing prokaryotes. Mineral biooxidation takes place in highly aerated, continuous-flow, stirred-tank reactors or in irrigated dump or heap reactors, both of which provide an open, non-sterile environment. Continuous-flow, stirred tanks are characterized by homogeneous and constant growth conditions where the selection is for rapid growth, and consequently tank consortia tend to be dominated by two or three species of micro-organisms. In contrast, heap reactors provide highly heterogeneous growth environments that change with the age of the heap, and these tend to be colonized by a much greater variety of micro-organisms. Heap micro-organisms grow as biofilms that are not subject to washout and the major challenge is to provide sufficient biodiversity for optimum performance throughout the life of a heap. This review discusses theoretical and pragmatic aspects of assembling microbial consortia to process different mineral ores and concentrates, and the challenges for using constructed consortia in non-sterile industrial-scale operations.
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              Structure and function of the molecular chaperone Trigger Factor.

              Newly synthesized proteins often require the assistance of molecular chaperones to efficiently fold into functional three-dimensional structures. At first, ribosome-associated chaperones guide the initial folding steps and protect growing polypeptide chains from misfolding and aggregation. After that folding into the native structure may occur spontaneously or require support by additional chaperones which do not bind to the ribosome such as DnaK and GroEL. Here we review the current knowledge on the best-characterized ribosome-associated chaperone at present, the Escherichia coli Trigger Factor. We describe recent progress on structural and dynamic aspects of Trigger Factor's interactions with the ribosome and substrates and discuss how these interactions affect co-translational protein folding. In addition, we discuss the newly proposed ribosome-independent function of Trigger Factor as assembly factor of multi-subunit protein complexes. Finally, we cover the functional cooperation between Trigger Factor, DnaK and GroEL in folding of cytosolic proteins and the interplay between Trigger Factor and other ribosome-associated factors acting in enzymatic processing and translocation of nascent polypeptide chains.
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                Author and article information

                Journal
                Microorganisms
                Microorganisms
                microorganisms
                Microorganisms
                MDPI
                2076-2607
                19 July 2020
                July 2020
                : 8
                : 7
                : 1076
                Affiliations
                [1 ]Winogradsky Institute of Microbiology, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Ave., 33, bld. 2, Moscow 119071, Russia
                [2 ]Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Malaya Pirogovskaya, 1a, Moscow 119435, Russia; d.matyushkina@ 123456gmail.com (D.M.); nikitishena@ 123456mail.ru (O.P.)
                Author notes
                [* ]Correspondence: zhuravleva-inmi@ 123456mail.ru ; Tel.: +7-499-135-65-96
                Author information
                https://orcid.org/0000-0002-2684-2287
                https://orcid.org/0000-0001-9535-5792
                Article
                microorganisms-08-01076
                10.3390/microorganisms8071076
                7409299
                32707712
                50a1bb9b-a4de-4a42-b17e-65b23ea03163
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 11 June 2020
                : 17 July 2020
                Categories
                Article

                acidophiles,sulfobacillus thermotolerans,sulfide concentrate,arsenic,resistance,differential proteomics,cellular element content

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